Repair of spinal cord injury by using the tissue engineered nerve grafts constructed by silk fibroin/filament including seed cells

doi:10.16098/j.issn.0529-1356.2015.05.004

Abstract

Abstract:

Objective To explore a method for construction of tissue engineered nerve grafts (tissue-engineered nerve grafts, TENGs) using silk fibroin/filament in vitro and to evaluate their roles for repairing spinal cord injury. Methods Isolation and induction of rat skin derived precursors into Schwann cells (SCs). S-100 immunofluorescence histochemical staining was applied for the SCs identification. As seed cells, SCs differentiated from skin derived precursors (SKPs) cultured with silk fibroin conduit and silk filament scaffold in vitro. After culturing for 7 days, the TENGs were transplanted into the injury site of rat dorsal spinal cord (T8-T10) hemisection model. BBB score was evaluated at different time points after the operation. The spinal cord samples were obtained 8 weeks after the operation and immunofluorescence histochemical staining was applied to evaluate the regeneration of the injured spinal cord and the survival of seed cells. Results Under a phase contrast microscope, most of the seed cells were bipolar or tripolar. The immunofluorescence staining showed that SKP-SCs were S-100 positive. After the transplantation of the TENGs into spinal cord injury site, BBB scores were significantly higher in TENGs group than in control group from 4 weeks to 8 weeks. The GFP-positive seed cells were also investigated after 8 weeks after the operation. Conclusion The TENGs constructed by silk fibroin/filament including seed cells may promote repairing of rat spinal cord injury.

Key words: Central nervous system, Tissue engineering, Schwann cell, Silk fibroin, Spinal cord injury, Immunofluorescence, Rat

LIU Xiao-dong XUE Cheng-bin JU Qian-qian YANG Xiao-hua QIN Jian-bing TIAN Mei-ling Lü Guang-ming*. Repair of spinal cord injury by using the tissue engineered nerve grafts constructed by silk fibroin/filament including seed cells[J]. AAS, 2015, 46(5): 596-601.

References

［1］Toma JG, Akhavan M, Fernandes KJ, et al. Isolation of multipotent adult stem cells from the dermis of mammalian skin［J］. Nat Cell Biol, 2001, 3(9): 778-784. 
［2］Yui S, Fujita N, Chung CS, et al. Olfactory ensheathing cells (OECs) degrade neurocan in injured spinal cord by secreting matrix metalloproteinase-2 in a rat contusion model［J］. Jap J Vet Res, 2014, 62(4): 151-162.
［3］Lee TH. Functional effect of mouse embryonic stem cell implantation after spinal cord injury［J］. J Exerc Rehabil, 2013, 9(2): 230-233. 
［4］Hong JY, Lee SH, Lee SC, et al. Therapeutic potential of induced neural stem cells for spinal cord injury［J］. J Biol Chem, 2014, 289(47): 32512-32525. 
［5］Richardson PM, McGuinness UM, Aguayo AJ. Axons from CNS neurons regenerate into PNS grafts［J］. Nature, 1980, 284(5753): 264-265.
［6］Azanchi R, Bernal G, Gupta R, et al. Combined demyelination plus Schwann cell transplantation therapy increases spread of cells and axonal regeneration following contusion injury［J］. J Neurotrauma, 2004, 21(6): 775-788. 
［7］Ding P, Yang Z, Wang W, et al. Transplantation of bone marrow stromal cells enhances infiltration and survival of CNP and Schwann cells to promote axonal sprouting following complete transection of spinal cord in adult rats［J］. Am J Transl Res, 2014, 6(3): 224-235. 
［8］Kanno H, Pearse DD, Ozawa H, et al. Schwann cell transplantation for spinal cord injury repair: its significant therapeutic potential and prospectus［J］. Rev Neurosci, 2015, 26(2): 121-128. 
［9］Biernaskie J, Sparling JS, Liu J, et al. Skinderived precursors generate myelinating Schwann cells that promote remyelination and functional recovery after contusion spinal cord injury［J］. J Neurosci, 2007, 27(36): 9545-9559.
［10］Khuong HT, Kumar R, Senjaya F, et al. Skin derived precursor Schwann cells improve behavioral recovery for acute and delayed nerve repair［J］. Exp Neurol, 2014, 254:168-179.
［11］Zhang N, Yan H, Wen X. Tissue-engineering approaches for axonal guidance［J］. Brain Res Brain Res Rev, 2005, 49(1): 48-64.
［12］Bellocchio EE, Reimer RJ, Fremeau RT Jr, et al. Uptake of glutamate into synaptic vesicles by an inorganic phosphate transporter［J］. Science, 2000, 289(5481): 957-960.
［13］Takamori S, Rhee JS, Rosenmund C, et al. Identification of a vesicular glutamate transporter that defines a glutamatergic phenotype in neurons［J］. Nature, 2000, 407(6801):189-194.
［14］Vigneault ?, Poirel O, Riad M, et al. Distribution of vesicular glutamate transporters in the human brain［J］. Front Neuroanat, 2015, 9:23.
［15］Brandt J, Evans JT, Mildenhall T, et al. Delaying the onset of treadmill exercise following peripheral nerve injury has different effects on axon regeneration and motoneuron synaptic plasticity［J］. J Neurophysiol, 2015, 113(7): 2390-2399.

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